Analyzing Cancer Cell Viability and Apoptosis

Cell Counting Kit-8 kit was purchased from DOJINDO (CK04, Kumamoto, Japan). Cancer cells (5000) or PBMCs (50,000) were placed on 96-well plates (100 μl/well). The experiments were performed as previously described [21 (link)]. After co-culture of cancer cells with PBMCs for 48 h, cells were harvested and stained with the antibodies labeled by fluorescence for 30 min in 4 °C in darks. Propidium Iodide (PI) (556463, BD, New Jersey, USA), FITC Annexin V (556420, BD, New Jersey, USA) were used. Fluorescence intensity was assayed by FACS Ariar (BD, New Jersey, USA), and analyzed using FlowJo 7.6.1 software (FlowJo, RRID:SCR_008520).